GB/T 36389-2018T4 DNA ligase (English PDF)
T4 DNA连接酶
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Issued by
SAMR; SAC
Level / Type
National · Recommended
Issue date
June 7, 2018
Implementation date
January 1, 2019
Scope
GB/T 36389-2018 is the English-translated version of T4 DNA连接酶.
GB/T 36389-2018 covers T4 DNA ligase isolated and purified from recombinant genetically engineered bacteria, the tool enzyme that joins adjacent DNA strands by a phosphodiester bond between the 5'-P and 3'-OH ends, linking sticky ends or blunt ends.
It specifies the technical requirements, test methods, packaging, transport, storage and shelf life of the enzyme. The requirements include a clear liquid without precipitation, an activity of at least 8000 U/mL and the absence of exonuclease and endonuclease.
The standard also defines the unit of activity through the ligation of HindIII-digested lambda DNA fragments in 30 min at 16 °C, and refers to YY/T 0087 electrophoresis devices and YY/T 0657 medical centrifuges. It was issued on 7 June 2018 and has applied since 1 January 2019.
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Document preview — GB/T 36389-2018
National Standard of the People's Republic of China
- ICS
- 07.080
- Classification
- C 27
Issued by: State Administration for Market Regulation; Standardization Administration of the PRC
Contents
- Foreword
- Introduction
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Technical requirements
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
Introduction
T4 DNA ligase is derived from recombinant genetically engineered bacteria and catalyzes the phosphodiester of the 5'-P terminus and the 3'-OH terminus of adjacent DNA strands.
The bond-binding reaction promotes the connection between the viscous ends or between the blunt ends. Develop a national standard for T4 DNA ligase to promote this The industrialization of class tool enzymes is of great significance.
1 Scope
This standard specifies the technical requirements, testing methods, packaging, transportation, storage and shelf life of T4 DNA ligase.
This standard applies to T4 DNA ligase isolated and purified from recombinant genetic engineering bacteria.
2 Normative references
The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this document. For undated references, the latest edition (including all amendments) applies to this document.
GB/T 191 packaging storage and transportation icon
GB/T 6682 Analytical laboratory water specifications and test methods
YY/T 0087 electrophoresis device
YY/T 0657 medical centrifuge
3 Terms and definitions
The following terms and definitions apply to this document.
3.1 T4 DNA ligase
A nucleic acid ligase that catalyzes the reaction of the 5'-P terminal and the 3'-OH terminal of a neighboring DNA strand by a phosphodiester bond The connection between the sticky ends or between the flat ends.
3.2 Unit of activity
In 20 µL of 1×T4 DNA ligase reaction buffer, 50% of HindIII digestion can be achieved in 30 min under 16 °C reaction conditions.
The amount of enzyme required for ligation of the lambdaDNA fragment [5' end concentration of 0.12 µmol/L (300 µg/mL)] was defined as 1 unit of activity (U).
4 Technical requirements
4.1 Appearance
Clarify the clear liquid without precipitation.
4.2 T4DNA ligase activity
>=8000U/mL.
4.3 Impurities
Exonuclease and endonuclease should not be included.
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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.
Referenced standards
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Related Standards
GB/T 191-2025 — Graphical symbols marking for handling and storage of packages
GB/T 6682-2008 — Water for analytical laboratory use - Specification and test methods
YY/T 0087-2004 — Electrophoresis device
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